The impact of culture conditions on blastocyst formation and aneuploidy rates: a comparison between single-step and sequential media in a large academic practice



To compare a single-step medium with a sequential medium on human blastocyst development rates, aneuploidy rates, and clinical outcomes.


Retrospective cohort study of IVF cycles that used Sage advantage sequential medium (n = 347) and uninterrupted Sage 1-step medium (n = 519) from July 1, 2016, to December 31, 2017, in an academic fertility center. Main outcome measures are blastocyst formation rates per two-pronuclear (2PN) oocyte and aneuploidy rates per biopsy.


Of all IVF cycles, single-step medium yielded higher blastocyst formation rate (51.7% vs 43.4%) but higher aneuploidy rate (54.0% vs 45.8%) compared with sequential medium. When stratified by maternal age, women under age 38 had no difference in blastocyst formation (52.2% vs 50.2%) but a higher aneuploidy rate (44.5% vs 36.4%) resulting in a lower number of euploid blastocysts per cycle (2.6 vs 3.3) when using single-step medium compared to sequential medium. In cycles used single-step medium, patients ≥ age 38 had higher blastocyst rate (48.0% vs 33.6%), but no difference in aneuploidy rate (68.8% vs 66.0%) or number of euploid embryos (0.8 vs 1.1). For patients reaching euploid embryo transfer, there was no difference in clinical pregnancy rates, miscarriage rates, or live birth rates between two culture media systems.


Our study demonstrates an increase in aneuploidy in young women whose embryos were cultured in a single-step medium compared to sequential medium. This study highlights the importance of culture conditions on embryo ploidy and the need to stratify by patient age when examining the impact of culture conditions on overall cycle potential.

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The authors thank the staffs at Stanford IVF center for their clinical support of the study.

Authors’ roles

J.D. and Q.Z. were involved with study design, execution, and statistical analysis. J.D. wrote the manuscript. C.C. and R.K. coordinated data collection and statistical analysis. R.L. contributed interpretation of the data and edit of the manuscript. B.B. devised, supervised the study and edited the manuscript. All authors reviewed the manuscript and provided critical feedback and discussion.

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Correspondence to Barry Behr.

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Conflict of interest

Both Cengiz Cinnioglu and Refik Kayali disclosed that they are employees of Igenomix Inc. All other authors do not have conflicts of interest to disclose.

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The work was done in Stanford Medicine Fertility and Reproductive Health Services.

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Deng, J., Zhao, Q., Cinnioglu, C. et al. The impact of culture conditions on blastocyst formation and aneuploidy rates: a comparison between single-step and sequential media in a large academic practice. J Assist Reprod Genet (2020).

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  • Culture media
  • Embryo development
  • Aneuploidy
  • Infertility
  • Pregnancy